DD159908A5 - FUEL MATERIAL FOR QUICK-FLUID CHROMATOGRAPHY AND METHOD FOR THE PRODUCTION THEREOF - Google Patents

Abstract

A granular crosslinked copolymer useful as a filling material for high speed liquid chromatography and, more particularly, as a gel permeation chromatography filling material is explained. In a preferred embodiment of the invention, the filler material is a copolymer consisting essentially of (I) units of at least one vinyl alcohol, (II) units of at least one vinyl ester of a carboxylic acid and (III) units of at least one crosslinking monomer having an isocyanurate ring, wherein the ratio of the units (I) to the units (II) in the copolymer is within the range corresponding to the following equation: (a) wherein a and b are molar ratios of the units (I) and (II) in the total units ( I), (II) and (III) of the copolymer, and a process for its preparation by suspension polymerization will be described.

Description

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Filling material for high-speed liquid chromatography and process for its preparation

Application of the invention;

The invention relates to a novel granular crosslinked copolymer, a filling material for the fast liquid chromatography (abbreviated "HLC) consisting of this crosslinked Copolytneres, and a method for its preparation, More specifically, the invention relates to a filler for di <liquid chromatography (abbreviated" LC ") which is suitable for a rapid and high-grade separation or analysis of substances dissolved in an aqueous solution according to the separation mechanism which is practically prevalent in gel permeation chromatography (abbreviated "GPC") and which has a copolymer consisting essentially of (i) units of at least one vinyl alcohol , (II) units of at least one vinyl ester of a carboric acid and (III) units of at least one crosslinking monomer <with an isocyanurate ring, and a process for the <preparation.

Characteristic of the known technical solutions:

The LC is a method of separation or analysis, in which the differences in the Eluierungsgeschvvindigkeit be used by some interaction between the solid phase, d _# h. a filler material, and to be separated, be dissolved in the movable component be movable liquid phase components. Above all, a method in which a filler having a small particle size and high mechanical strength is applied, and a high degree of separation or analysis is achieved by passing a solvent at high speed in a short time generally

referred to as HLC and used in various fields «

The GPC is a type of LC which employs the principle that a component having a smaller molecular size than the pore size in the filler (referred to as "gel") will enter the gel due to its molecular size, while passes through a Koraponente with a larger molecular size outside the gel, whereby the components can be successively eluted with a solvent in the order of the component with the larger molecular size.

The GPC may be after for separation or analysis? used solvents are divided into an organic medium system and an aqueous medium system. Of these two systems, the aqueous medium GPC system can be used for the separation or analysis of water-soluble synthetic polymers, saccharides, amino acids and proteins. It deserves especially attention in biochemical and medical fields as a simple analytical method that the analysis can be applied, for example, serum advantageous för without the sample pretreated or the solvent must be changed during the analysis as ea in the conventional LC by applying of adsorption and distribution is the case, and it also provides a greater amount of information. In particular, the components in the blood and urine are said to be closely related to kidney or liver diseases or to symptoms such as carcinoma. Thus, it is urgently needed to develop gels for the HCL, especially gels for a fast GPC system with aqueous medium suitable for the separation or analysis of these components. There is a gel and will

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frequently used as the gel for an aqueous medium GPC system _/ produced by the crosslinking of dextran with epichlorohydrin (trademark "Sephadex", a product of Pharmacia Fine Chemicals AB, Schvyeden). However, this gel is a so-called soft gel whose pores used for the separation consist of crosslinked networks, but the mechanical strength of the gel is low. Therefore, this gel could not be used as a gel for the fast GPC.

It is also known that a gel in the form of an aqueous medium can be obtained by saponification of the copolymer particles of vinyl acetate and 1,4-butanediol divinyl ether, as set forth in U.S. Patent 3,586,626. However, as added by W, Heitz, the inventor of this U.S. patent, the copolymerizability of the monomers used for this gel is low (see IV, Heitz, O, Chromatogr, 53, 37, 1970), and therefore the gel formed has no sufficient strength and practically can not be used for the HLC.

It was also reported that a gel in the form of an aqueous medium of the saponified copolymers can be produced with a higher mechanical strength by saponification of the copolymer of example Diäthylenglycoldimethacrylat or diglycidyl methacrylate with vinyl acetate and subsequent crosslinking, as set forth in U.S. Patent No. 4,104,208. However, such a production method is complicated, and it is difficult to obtain the gel in a consistent quality with good reproducibility.

It is also known that a hard polyvinyl alcohol gel

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can be obtained when a copolymer of vinyl acetate and a crosslinking agent with a triazine ring structure are saponified to a degree that the ester absorption of the infrared absorption spectrum at 1730 cm completely disappears, as described in GB "PS 2034328 A. According to the invention underlying research, however, the gel obtained by this method has a low mechanical strength, and therefore, the gel does not for fast-GPC is suitable for aer the gel with a smaller particle size, e.g., B, a Geivichtsmittel-Teilchendurchniesser is used below about 20 mM under high pressure, even though the gel may find use in the größtechnischen separation in which the gel can be used with a higher particle size and no particularly high mechanical strength is required as in the desalination of an aqueous Polynierlösung _e

So that the gel is suitable for a Schnei1 GPC Systetn with an aqueous medium, there must have pores which are accurately controlled according to the molecular sizes of 6 components to be separated, and it has a small particle size have a sufficient mechanical strength, and besides, it must hydrophilic with low adsorbability the components to be separated in an aqueous solution

In view of the prior art described above, extensive studies have been made for the development of gels for an aqueous medium GPC system which satisfy the above-mentioned requirements. A gel of predominantly hydroxyl groups, ester groups and units of a crosslinking monomer in the polyraer skeleton with pores used for the separation or analysis of corresponding substances in a poly

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ner aqueous solution, and a mechanical strength that endures high speeds or high pressures and discovered a method for the production of the same.

Darlequ njg_des essence he d Erf indung;

In one embodiment, the invention provides a fully porous fast liquid chromatography filler comprising a granular cross-linked copolymer consisting essentially of (I) units of at least one vinyl alcohol, (II) units of at least one vinyl ester of a carboxylic acid, and (III) Units of at least one crosslinking monomer with an isocyanurate ring, the ratio between aen units (I) and the units (II) in the copolymer being in the range corresponding to the following equation:

about 0.4 = = about 0.8 where

yes and b molar ratios of the units (I) or

(II) throughout the units (I), (II) and

(III) of the copolymer are «

In another embodiment, the invention provides a fully porous fast liquid chromatography filler comprising a granular crosslinked copolymer consisting essentially of (I) units of at least one vinyl alcohol, (II) units of at least one vinyl ester of a carboxylic acid and (III) units of at least one crosslinking monomer having an isocyanurate ring, wherein the ratio of units (I), (II) and (III) in the copolymer is in the range corresponding to the following equation:

about 0.20 "~ 7 ~ V-3c = about 0.40 where

£ and t) have the same meaning as defined above, and £ is the molar ratio of units (HI) in the total units (I), (II) and (III) of the copolymer.

In another embodiment, according to the invention a process for the production of the above-described fullage materials is made available

In the drawings:

Figure _e 1 is a calibration curve for an inventive GPC-gel with the definition of Mu _n, / *

Fig. 2 is an infrared absorption spectrum of the gel of Example 1;

Fig. 3 to 5 chromatograms, which were obtained using the gels of Examples 1, 2 and 5.

The gel for use in a Schneil-GPO system with aqueous medium must have a mechanical strength which can withstand high speeds or high pressures and hydrophilicity. The hydrophilicity of the gel according to the invention is based on the hydroxyl groups present in the polymer skeleton. Such hydroxyl groups may be prepared by the Esteraustausch. or saponification of the ester groups in a copolymer of at least one vinyl ester of a carboxylic acid and at least one crosslinking monomer having an isocyanurate ring. However, the investigations according to the invention have shown that the mechanical strength of a

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Gels is reduced if all ester groups ester-exchanged or saponified. If too many unchanged ester groups are present, the hydrophilicity of the gel is reduced and the components dissolved in an aqueous solution become too readily adsorbed. It has now been found convenient to carry out the ester interchange reaction or saponification to such an extent that about 40 to about 80 percent of the ester groups are converted to hydroxyl groups. The Esteraustauschverhältnis or Verseifungsverhältnie Y (O ^ Y $ 1) is represented by the following Tung DC (1):

3 and Jb are molar ratios of (I) the units of at least one vinyl alcohol and (II) the units of at least one vinyl ester of a carboxylic acid in the copolymer,

that is, Y denotes the ratio of units (I) to units (II) in the copolymer.

a. and Jb can be calculated from the density of the hydroxyl groups (referred to as "q _0H ") in the gel and the amount of (III) units of at least one crosslinking monomer having an isocyanurate ring. q _0H is the equivalent of the units (I) per unit mass of the gel and can be determined by reacting the gel with acetic anhydride in a pyridine solvent by measuring the amount of acetic anhydride consumed in the reaction with the hydroxyl groups or the mass change of the gels and calculating the concentration of hydroxyl groups from this measured value For example, when 1 millimole of acetic anhydride is consumed in the reaction with 1 g of a dry gel, the q _{QH of} this gel is 1 meq / g

14 f% Γ * * 7 105

.the units of crosslinking monomers having an isocyanurate can be identified by its infrared absorption spectrum, and its amount can be determined from the nitrogen content, which is determined by the Eleraentaranalyse the gel * In other words, £ may consist q _QH and t> from dera Value obtained by subtracting the total amount of units (I) and (III) from the total amount of units (I), (II) or (III) in the gel, respectively. The chemical structure of the vinyl carboxylate groups can be confirmed by identifying the carboxylic acid formed by the complete ester exchange reaction or saponification of the gel. If the conditions for the preparation of the gel are known, it is also possible to calculate Y from the composition of the starting materials and q _0. , From the gel formed. It is preferred that the Esteraustauschverhältnis or the saponification in the range from about 0.45 to about 0.75 should be "Even if q _Q., Depending on the used as AusgangsBiaterial Vinylester of the carboxylic acid, the degree of crosslinking and the Esteraustauschverhältnis or dera saponification varies, it is according to the invention is generally 3 to 11 meq / g _e this has the inventive gel having the respective units of the vinyl alcohols, the Vinylesters the carboxylic acid and the crosslinking mono Raeren having an isocyanurate ring at the same time, excellent characteristics as a gel for the quick- -GPC "This could be due to the fact that the units of residual ester groups in the gel contribute to maintaining the mechanical strength of the gel, and also to the fact that these units have a higher hydrophilicity than the units of the crosslinking monomer, thus rendering the adsorbent can be reduced as in the case where the same mechanical strength is maintained only dtarch the Vernetzungsmönoraere »·

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Among the gels prepared by ester exchange or saponification of a copolymer of the vinyl ester of a carboxylic acid and the crosslinking monomer having an isocyanurate ring, the gel prepared by using a larger amount of the crosslinking monomer has higher mechanical strength. However, since the crosslinking monomer does not contain a hydroxyl group, and therefore, no hydroxyl group can be formed by hydrolysis, the gel of a copolymer containing a larger amount of crosslinking monomer units in the polymer skeleton has a low hydrophilicity. In order for the gel to be useful for the fast GPC, the gel must contain the isocyanurate units within an optimal range. It is beneficial if the degree of crosslinking (referred to as "X") is in the range of about 0.2 «X = about 0.4. X is represented by the following formula (2):

^{X s}

a + b + 3c wherein

£ and Jj are as defined above and £ is the molar ratio of units (III) described above in units (I), (II) and (III) of the copolymer.

In the above formula, yes and t> can be determined from the values of elemental analysis of the gel or of the esterified gel by the method described above and £. When the conditions for the preparation of the gel are known, X can be easily determined by calculating the molecules of the vinyl ester of the carboxylic acid and the crosslinking monomer used for the polymerization, which are substituted for (a + b) and ε, respectively.

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- ίο -

If X in the above-specified range, the gel will have a sufficient strength with a small particle size and aen use under conditions of high pressure and high speed withstand "Such a gel is sufficiently hydrophilic and the components in an aqueous solution, especially proteins and amino acids "absorb difficult ad · what is a big advantage for use in the fast-GPC _e Moreover, it is particularly advantageous if the inventive gel at the same time meets the above specified ranges of X and Y.

In the conventional soft gel, to increase an exclusive molecular weight (which is called "Mi"), d _e g. "network are slightly reduced, which inevitably leads to the recovery of water (referred to as" a minimum molecular weight of a substance which can not penetrate into the pores of the gel, uer degree of crosslinking of the extension of the referred W _R ") to give Particularly with a small particle size of the gel, unfavorable effects such as pressure loss in a packed column may occur due to reduced mechanical strength. Therefore, gels having a particle diameter of at least 50 AJm are generally used.

In contrast, the gel of the present invention has a W "of from about 0.5 to about 2.0 grams of water / g dry gel, independent of Μ ,. , and therefore even the gel with high M ,. used for the fast GPC. This is especially true for gels in the form of an aqueous medium for rapid GPC in which gels having a particle diameter of at most 20 Aim and which must have sufficient mechanical strength can be used. W _R is defined as the amount (g) of water that may contain one gram of dried gel, if

- Il -

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the gel has been equilibrated with water and it is a measure of the amount of pores in the gel that perform the GPC function. At elevated W _R , the mass of polymer ketone moieties per unit volume of the gel in water, ie, the mass of the gel as such, is correspondingly reduced. Therefore, when W _{R is} too high, the mechanical strength of the gel in water is reduced, and it is impossible to increase the flow rate, resulting in a pressure loss in the column. On the other hand, when W ~ is too small, the amount of pores in the gel having the GPC function is lowered, so that the releasability of the gel is reduced. Thus, in the case of a gel in the form of an aqueous medium for rapid GPC, it is critical to achieve a W _R within a reasonable range.

It is convenient if the gel for quick GPC having a structure according to the invention has a W _{R of} from about 0.5 to about 2.0 g water / g dry gel, especially from about 0.8 to about 2.0 g water / g dry gel for practical purposes. The gel of the invention may have a W _R within such a range. W ..? is determined as follows:.

A gel immersed in water in a perfectly equilibrated state is centrifuged to remove water attached to the surface of the gel, and its mass (W ₅ ) is determined. The gel is dried and its dry weight (W ₂ ) is determined, W _R is calculated using the following formula:

- W

~ 12 -

31 105 7

The M of the gel of the present invention may vary within a wide range. As described above, Μ · bezeichnet denotes the relative minimum molecular weight of a substance that can not penetrate into the pores of the gel. Substances with a molecular weight below this critical value can be separated by GPC, but substances with a relative molecular mass above this critical value can not penetrate into the pores of the gel, but pass directly through interstices between the gels The latter substances have substantially the same elution volumes regardless of the molecular weight, and therefore such substances can not be separated by GPC.

M ,. can be determined from the GPCE calibration curve. The GPC calibration curve can be obtained by plotting the logarithm of the molecular weights of individual samples whose molecular weights are known, on the ordinate and the elution volume of each sample on the abscissa with respect to a column filled with gel, as shown in FIG , It represents the relationship between the elution volume and the relative molecular weight of a material to be separated in the chromium * togram. The inclined line and the ordinate parallel line in this diagram are substantially straight, and the section where the two straight lines meet is a curve. M ,. is expressed as the value on the ordinate at a point at which the extension of the inclined line intersects the extension of the line parallel to the y-ordinate, M.- ·· is a property typical of the gel and represents the exclusive Relative molecular pathway in which the gel can exert a release effect due to differences in the size of the molecules. Substances having a molecular weight greater than the exclusive molecular weight are substantially eluted together without separation.

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According to the invention M _lim is determined so that polyethylene glycols or dextranes and distilled water are used as the solvent as a standard substance having a known molecular weight. Since commercially available water-soluble standard polymers have less than about 2,000,000 relative molecular weight, it is sometimes not possible to obtain a complete calibration curve for a gel with more than 2,000,000 M ,. set up. Therefore, the Mj. of such a gel is not accurately determined, but is intersected by the point of intersection at which the extension of the relative molecular weights of less than 2,000,000 relative molecular masses intersect the extension of the line parallel to the y-ordinate with respect to a gel lower M ,. was determined under the same conditions.

In order for the gel to have a mechanical strength suitable for the fast GPC and to have no adsorbability in combination, the degree of crosslinking X is preferably in the range of about 0.24 to about 0.32 when M ,. is in the range of about 10 to about 10, and the degree of crosslinking X j is preferably in the range of about 0.27 to about 0.35 when M ,. ira range from about 10 to about 10 «

In addition, the gel of the invention, which is a completely porous hard gel, has a large specific effective surface in the dry state. The completely porous structure is understood here to mean a structure which has fine pores evenly distributed throughout the inner regions of the particles. Generally, it can be seen that organic synthetic polymers having a crosslinked structure in a solvent with affinity for polymers will swell and shrink on drying. In the case of a soft gel, at the time of the

Swelling with a solvent-filled pore can only be maintained by the crosslinked network when the gel is dried, such a network can maintain the swollen state and therefore will shrink, and the pores will largely disappear. In such a case, the specific effective surface area will substantially take the value at the outer surface of the particles and will generally show a very low value of less than 1 m / g *. On the other hand, in the case of a hard gel having a solid porous structure, the pores may the gel was dried to maintain the same state as it had at the time of swelling, albeit with slight shrinkage. In other words, the pores have a permanent texture. Therefore, the hard gel has a far greater specific effective surface area than the soft gel.

The gel of the invention generally has a specific effective surface area of from about 5 to about 1000 m / g. If the gel has a specific effective surface area lower than the lower limit of the range, it means that the gel has a kind of homogeneous structure (soft gel) and hardly contains fine pores and hence is not suitable for the fast GPC.

The specific effective surface area can be measured by various conventional methods, but for the invention it is measured by the most commonly used BET method using nitrogen gas. In addition, the samples used to measure the specific effective surface area must be sufficiently dry. Since the gel of the invention is difficult to dry because of the high hydrophilicity, it is preferred to first equilibrate the wet gel with acetone and then under reduced pressure at room temperature to dry below 60 ⁰ C temperature.

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The gel of the invention generally has a weight average particle diameter (referred to as "D ₁₁ ")

from about 4 to about 200 / um. Particularly excellent features are exhibited by the gel for HLC when δ is low /, for example, in the range of about 5 to about 20 μm. If a particularly high release capability is required, then it is beneficial if the gel has a D _w of about 5 to about 12 μm. D, will with the help of a coulter

/ w

Counters (a product of Coulter Electronics, Inc _{# /} USA) and calculated according to the formula:

0 - ^ Cni. di ⁴ ) ^w " Z (ni, di ³ ) where

di is the particle diameter, and ni is the frequency of occurrence of particle diameters

It is well known in the field of liquid chromatography that the ability to separate can be improved by choosing a smaller particle size of a filler. However, when a solvent is passed through a column filled with the smaller particle size gel, the pressure loss is greatly increased as compared with a case where a gel having a larger particle size is used. Therefore, when the gel has a low mechanical strength, it is deformed or broken, resulting in an excessively high pressure loss, which makes HLC impossible by using a gel having a small particle size. Since the mechanical strength of the gel of the invention can be successfully improved by controlling various properties, for example, the ester exchange ratio or the saponification ratio and the degree of crosslinking, the gel can be used despite the

small particle size withstand high speeds and high pressures »

The gel of the present invention having the above-described properties can be produced by sparingly dissolving in water a suspension polymerization of monomer gas of at least one vinyl ester of a carboxylic acid and at least one crosslinking monomer with an isocyanurate ring in the presence of a diluent capable of dissolving the monomer , is carried out, and ester interchange or Verisi "tion of the resulting copolymer particles is carried out to an extent that the ester exchange ratio or the saponification ratio between about 0.4 and about 0.8.

The vinyl ester of carboxylic acid usable for the invention is a compound having at least one polymerizable vinyl carboxylate group which can be selected from vinyl esters of a carboxylic acid having 2 to 5 carbon atoms. Examples of such vinyl esters of the carboxylic acid include vinyl acetate, vinyl propionate, vinyl butyrate, vinyl valerate and the like Vinyl pivalate, which can be used singly or as a combination of two or more species. Of these vinyl esters of carboxylic acid, vinyl acetate and vinyl propionate are particularly preferred because of easy polymerization / light ester exchange, easy saponification, and continuous availability.

The crosslinking monomer having an isocyanurate ring usable for the invention is represented by the following formula:

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R ₁ . σ R ₂

NN

ο · ο

V

R ₁ , R ₂ and R _{3 may be the} same or different and selected from the group consisting of -CH ₂ -CH = CH ₂ , -CH-C = CH and -CH ₂ -C = CH ₂ .

Of these crosslinking monomers, triallyl isocyanurate (R ₁ SsR ₂ = R ₃ S -CH ₂ -CH = CH ₂ ) has good copolymerizability with vinyl acetate and good ester exchange reaction or saponification resistance, and is therefore preferred as a crosslinking monomer.

The molar ratio of the vinyl ester of the carboxylic acid and the employable crosslinking monomer is generally 1 to about O / O8 to about 0.22. According to the use of the recovered gel, the molar ratio of the vinyl ester of the carboxylic acid to the crosslinking monomer used is preferably 1 to about 0.11 to about 0.16, or preferably 1 to about 0.12 to about 0.18.

Other monomers, such as 2-ethylhexyl methacrylate and acrylamide, which are copolymerizable with the vinyl ester of the carboxylic acid and the crosslinking monomer may also be used in an amount of, for example, up to 3% by weight relative to the mass of the vinyl ester of the carboxylic acid, where Properties and the Trenhleistung of the gel are not significantly adversely affected.

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Thus, continuous pores are formed in the Copolyraerteilchen and simultaneously the amount of AEC pores, the pore size and the size distribution of the pores can be controlled suspension polymerization system is at least one diluent is added, which can dissolve the monomer mixture but is hardly soluble in water, ,

Examples of such diluents usable in the present invention include aromatic hydrocarbons such as toluene and xylene; aliphatic hydrocarbons such as heptane, octane, decane and decalin; Ester compounds such as n-butyl acetate, isobutyl acetate and n-hexyl acetate; methyl ethyl ketone; and n-heptanol, which may be used singly or as a combination of two or more species. "A suitable amount of the diluent for use in the present invention is from about 20 to about 200 parts by weight per 100 parts by weight of the monomeric mixture. *** If the amount of diluent is below the lower limit If it is above the upper limit of the range specified above, the mechanical strength of the gel is reduced and the gel becomes suitable for use with small particle diameters unsuitable at high pressures or high speeds. "A preferred amount of diluent is in the range of about 20 to about 100 parts by weight per 100 parts by weight of monomer mixture.

In addition, to control pore size and pore size distribution of the gel, a linear polymer which is soluble in the monomer mixture can be used in conjunction with the diluent described above. "

In the linear used according to the invention. Polymer acts

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It is a linear polymer that may be soluble in the monomer mixture at a concentration of at least one mass% , and examples of such linear polymers are polyvinyl acetate and polystyrene. The amount of the linear polymer used is generally up to about 10 parts by weight, preferably about 0.3 to about 10 parts by weight per 100 parts by weight of the monomer mixture. By using the linear polymer in conjunction with the diluent, a gel can be easily generated which has a larger pore diameter, ό _φ h »a high M ,. , ,

Any normally used radical polymerization initiator may be used as the initiator for the suspension polymerization of the present invention. Examples of radical polymerization initiators include azo type initiators such as 2/2'-azobisisobutyronitrile and 2,2'-azobis (2,4-dimethylvaleronitrile and peroxide type initiators such as benzoyl peroxide , Lauroyl peroxide, di-t-butyl peroxide and cumene hydroperoxide. The amount of radical polymerization initiator is generally between about 0.1 and about 5 parts by weight per 100 parts by weight of the monomer mixture.

In carrying out the suspension polymerization, it is advantageous to add in the aqueous phase a suspension stabilizer which is selected from commonly known organic polymer stabilizers such as polyvinyl alcohol and methyl cellulose. The amount of suspension stabilizing agent used is generally about 0.1 to about 3% by weight relative to the mass of water used as the suspension polymerization medium. If necessary or desired, a pH buffering agent such as sodium phosphate may be used in combination. It is favorable if the pH of the suspension solution is kept approximately neutral. The particle size of the obtained by the polymerization

 - 20 -

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Granular copolymer can be varied by changing the type and amount of suspension stabilizer or stirring rate.

The volume or mass ratio between the organic phase and the aqueous phase is not very critical in the suspension polymerization, and it may be that normally chosen for carrying out the suspension polymerization.

The applied Polysnerisationsteraperatur is generally about 50 ⁰ C to about 90 ⁰ C, preferably about 60 ⁰ C to about 80 ⁰ C.

The applied polymerization time is generally between about 10 and about 50 hours,

After polymerization, the resulting Copolyinerteilchen are separated by filtration, washed thoroughly with water, hot water, "acetone, etc. _e to remove linear polymer Suspen · sion stabilizing agents, residual monomers, diluent etc _e, which adhere to the particles, and then dried.

The copolymer obtained in this way the Esteraustauschreaktion or saponification are subjected. The ester interchange reaction may be carried out in an alcohol such as methanol, ethanol, etc., as solvent, and the saponification may be carried out in water or a mixture of water and an alcohol such as methanol, ethanol, etc., as a solvent, each containing an acid such as sulfuric acid.

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acid, etc., or an alkali such as sodium hydroxide, potassium hydroxide, etc. is used.

The methodology used to Esteraustauschreaktion or the saponification temperature is generally between about OC and about 60 ⁰ C and preferably between about ⁰ ° C and about 40 ⁰ C _7, and the time for the Esteraustauschreaktion or the saponification is from about 5 to about 50 hours.

In order to produce a gel whose particle size is small and which can withstand high pressures or high rates, it is required in the invention that the ester exchange ratio or saponification ratio be in the range of about 0.4 to about 0.8. When all of the ester groups in the gel are ester-exchanged or saponified, ie, when the ester exchange ratio or saponification ratio is 1.0, a highly hydrophilic gel can be obtained, the mechanical strength of which is not sufficient in any case. A preferred ester exchange ratio or saponification ratio is about 0.45 to about 0.75. The conditions for the ester interchange reaction or the saponification may be appropriately selected by varying the kind of the reaction solvent, the reaction temperature, the reaction time, etc.

After the ester exchange reaction or saponification, the resulting particles are separated by filtration and washed thoroughly with water or hot water. If required, the particles are classified so they can be used as fill material for the HCL, by dispersing them in water, for example, to perform a simple classification utilizing the difference in the rate of sedimentation.

The gel of the present invention mainly contains hydroxyl groups, ester groups and units of the crosslinking monomer with the isocyahurate ring and has sufficient hydrophilicity at such a hydroxyl content within the above-mentioned range, and therefore, the gel shows no adsorbability to many solutes dissolved in water or analysis of water-soluble synthetic polymers, saccharides or proteins, the gel gives calibration curves in which the relationship between the elution volumetric properties and the logarithm of the molecular weight of a material to be separated can be represented by substantially straight or gentle curves. The gel of the present invention is thus usable as the GPC gel in the form of an aqueous medium. In the analysis of samples consisting of various kinds of components, such as serum or urine, in the use of the gel of the present invention, it is observed that some components are weak on the gel Were adsorbed on the gel, suggesting a higher elution volatility than that assumed by their relative occupants. As a result, a number of peaks are observed. In addition, the gel hardly exhibits any adsorbability of proteins such as albumin and elutes the proteins in a volume corresponding to their relative molecular mass. Therefore, in the analysis of serum or urine using the gel according to the invention, it is not necessary to remove proteins, and the process can be very In this case, the gel according to the invention can contain hydroxyl groups, ester groups and units of the crosslinking monomer with an isocyanurate ring in appropriate proportions

 "" '. * ♦ -

be assumed that the gel obtained a certain adsorbability in addition to the separating action, which is based on the differences in the size of the molecules, as intended by the invention, resulting in good separation as described above.

The gel according to the invention has, in addition to such a chemical

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Structure also has a very high mechanical strength with W "controlled in the range discussed above. Thus, despite its small particle size, the gel of the present invention can withstand high pressures and high speeds at least requires that it (1) contains pores in the gel, (2) has a low adsorbability, and (3) has such a high mechanical strength that it can withstand high pressures or high velocities even with small particle sizes meet these requirements, and these characteristic features can be achieved by meeting the above-specified ranges of X _{1 of} the ester exchange ratio or saponification ratio or W _R.

For example, such excellent characteristic separation characteristics and mechanical strength as described above can not be required of a gel having a barrier of a copolymer of vinyl alcohol and triallyl isocyanurate in which X is low and the infrared absorption spectrum at 1730 era completely disappears As explained in GB-PS 2034328 A, therefore, such a gel is not suitable as a filling material for the HCL.

The pore size of the gel of the present invention can be determined within a very wide range. Therefore, the gel of the present invention is useful not only for the separation or analysis of water-soluble synthetic polymers, saccharides or proteins, but also for the analysis of components having a molecular weight of several hundreds of thousands to several millions as present in blood and urine which are said to be closely related to kidney or liver diseases or to symptoms such as

r 24-

3 4 «Λ Γ FJ s 1 05 7

Since the gel according to the invention is also provided with excellent characteristic features such as a gel for the fast GPC as described above, these analyzes can be carried out within a short period of time and many information can also be obtained.

The gel according to the invention can generally be used in a state filled in a column. The column used is usually a stainless steel cylinder, but any other column may be chosen according to the intended purpose.

The invention will now be explained in more detail with reference to various examples, all of which are illustrative only and are not intended to limit the invention in any way.

example 1

A uniform mixture of 100 g of vinyl acetate, 32.3 g (X << 0.25) of triallyl isocyanurate, 100 g of n-butyl acetate and 3.3 g of 2,2'-azobisisobutyronitrile and 800 ml of an aqueous solution containing 1% by weight of a polyvinyl alcohol containing a degree of polymerization of about 2400 and a saponification ratio of 88 mole%, 0.05% by weight of sodium dihydrogen phosphate dihydrate and 1.5% by weight of disodium hydrogen phosphate dodecahydrate were charged to a 2 liter flask. After grundlichem mixing, the suspension polymerization was performed for 18 hours under stirring and heating to 65 ⁰ C and anschließend.5-hours at 75 ⁰ C. The resulting. Polymer particles were separated by filtration, with water and then with acetone

231105 7

washed, dried and then for 20 hours an ester exchange reaction in an out 47 g of sodium hydroxide and 2 1 methanol existing solution at 15 ⁰ C subjected. The resulting polymer particles were collected by filtration, washed with water and dispersed in water to carry out simple classification taking advantage of the difference in sedimentation velocity.

The weight average particle diameter (D ") of the thus-obtained crosslinked polyvinyl alcohol gel was measured by means of a Coulter Counter ZB (a product of Coulter Electroni Co., USA) and found to be 10.0 / μm The density of hydroxyl groups Cq ₀ Lj ) was determined by the method described above to give 7.3 meq / g, indicating an ester exchange ratio of 0.64 As shown in Fig. 2, after the ester exchange reaction by the infrared adsorption spectrum, it was also confirmed that ester groups remained in the polymer skeleton Infrared spectrum was measured by shaping the gel into KBr tablets and using an OASCO Infrared Spectrophotometer Type A320 (a product of Oapan Spectroscopic Co., Ltd., Oapan) The gel had a water recovery of 1.58 g Water / g dry gel and a specific effective surface area of 95 m / g.

This gel was filled in a stainless steel column of 7.5 mm in diameter and 50 cm in length, and aqueous solutions of dextrans and polyethylene glycols having different molecular weights were measured using as a detector a differential refractometer ("Shodex RI Model SE-II ", a product of Showa Denko Kabushiki Kaisha) was used. The substances of each group were consequently eluted in order of higher molecular weight, and it was demonstrated that

The separation was performed by the GPC. "In the analysis, the pump and the sample injector were those used by Hitachi Model 635A (a product of Hitachi Ltd ;, Oapan) and the detector of the Shodex RI / Model SE-II. The exclusive relative

4 molecular weight (M,.) Of dextran was about 3 χ 10. When the analysis of ^ -globulin, bovine serum-albumin / ovalbumin and myoglobin was carried out and as the solvent, an aqueous solution of 0.3 M sodium chloride and O _x IM sodium phosphate and an ultraviolet absorption detector ("Multi-Wavelength UV.Monitor", a product by Hitachi Ltd., Japan), the proteins were eluted in the order of the protein with a higher relative molecular mass and a recovery rate of substantially 100 % was achieved. All samples were measured at a flow rate of 1 ml / min

In addition, a sample solution of freeze-dried Huraanseruoi (trademark "ORTHO Normal Control Serum", a product of Ortho Diagnostics Inc ") was analyzed using the packed column to give a graph as in Fig. 3, in which the peak A is generally albumin, the peak B Creatinine and the peak C indicates uric acid. Some components were eluted because of their low adsorbability to the gel in an amount greater than the void volume of the column is, but a number of the components could · be detected and separated after elution of ^ globulin and albumin.

Example 2

A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 100 g of vinyl acetate, 32.2 g (X = 0.25) of triallyl isocyanurate, 40 g of toluene and 3.3 g of 2,2'- Azobisisobutyronitrile instead of the liquid

2 3110 5 7

mixed used in Example 1 and the ester exchange reaction. 20 hours at 40 ⁰ C performed.

The resulting gel had a D of 9.5 Aim, one of 9/0 meq / g (an ester exchange ratio of 0.74), a Wp of 1.0 g / g dry gel, and a specific active ingredient.

2 same surface of 38 ra / g.

This gel was filled in the same column as in Example 1, and the analysis of polyethylene glycols having different molecular weights was carried out using the packed column. As a result, it was confirmed that the elution order was with the polyethylene glycol having a higher molecular weight and M · · 1.9 × 10. All samples were reared at a flow rate of 1 ml / minute and each analysis completed within 20 minutes. In addition, a sample solution of freeze-dried human serum using the packed column was analyzed to obtain a graph as shown in Fig. 4, in which the peak D indicates albumin, the peak E creatinin and the peak F uric acid. Although the number of peaks appears to be lower than in Fig. 3 because of the lower sensitivity of the measurement, the basic course of the elution curve is almost the same as in Fig. 3. This confirmed that a number of components were detected and by the gel of this Example could be separated.

Example 3

A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 100 g of vinyl acetate, 37.5 g (X = 0.28) of triallyl isocyanurate, 100 g of n-butyl acetate and 4.1 g of polyvinyl acetate with a Polymerisierungs-

31 1 05

degree of about 500 and 3.4 g of 2 / 2'-AzObISiSObUtYrOnItTiI used instead of the liquid mixture of Example 1 and the ester exchange reaction carried out at 15 ⁰ C for 15 hours.

The resulting gel had an Mw of 9/1 , to, a of 5/1 meq / g (an ester exchange ratio of 0.50) / a Wp of 1/46 g of water / g dry gel and a specific effective

2 same surface of 86 m / g _e . ,

This gel was filled in the same column as in Example 1, and the analysis of polyethylene glycols and dextranes having different molecular weights and proteins such as α-globulin was carried out using the full-body column. It was confirmed that the substances of each group were in the order of Substance having a higher molecular weight were eluted and the proteins eluted at a recovery rate of substantially 100 %

were the M ₁ . for dextran 8 × 10, all samples were measured at a flow rate of 1 ml / minute and each analysis was performed within 20 minutes.

Example 4

A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 116 g of vinyl propionate / 39/4 g (X-0.29) triallyl isocyanurate / 62 g of n-butyl acetate and 3/9 g of 2/2 -Azobisisobutyronitrile used instead of the liquid mixture of Example 1 and the ester exchange reaction for 20 hours at 40 ⁰ C performed.

The resulting gel had a Dw of 10/8 Aim, one

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of 7.7 meq / g (an ester exchange ratio of 0.72), an IVp of 1.30 g of water / g dry gel and a specific active

same surface of 52 1117g,

This gel was filled in the same packed column as in Example 1, and the analysis of polyethylene glycols and dextranes having different molecular weights and proteins such as globulin was carried out using the packed body column. At this time, it was confirmed that the substances of each group were eluted in the order of the substance having a higher relative molecular weight, and that the proteins were eluted at a recovery rate of substantially % . for dextran was 2χ10. All samples were measured at a flow rate of 2 ml / minute and each analysis was completed within 10 minutes.

Example 5

A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 100 g of vinyl acetate, 41 g (X s 0.3) triallyl isocyanurate, 141 g of methyl isobutyl ketone and 2.8 g of 2,2'-azobisisobutyronitrile was used of the liquid mixture of Example 1 was used and the ester exchange reaction was carried out at 40 ^° C. for 20 hours.

The resulting gel had a Dw of 12.1 Aim, a q _0H of 7.8 meq / g (an ester exchange ratio of 0.62), a W _R of 1.78 g of water / g dry gel, and a specific active ingredient.

2 same surface of 87 m / g *.

This gel was placed in the same column as in Example 1.

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and the analysis of standard samples of polyethylene glycols and dextrans with different molecular weights was performed using the culture column. Substances of each group were eluted in the order of the substance with a higher relative molecular weight and the -Μ, of dextran was 5χ10. ♦ The analysis of standard samples of bovine serum burain, ovalburnin and myoglobin was also carried out using the packed column / $ 8 & these proteins were eluted in the order of the protein with a higher molecular weight and at a rate of recovery was essentially 100%. The resulting diagram is shown in Fig. 5 / in which the curve G is an elution curve of albumin / the curve H is an elution curve of ovalbumin and the curve I is an elution curve of myoglobin. As can be clearly seen from these results, the gel of this example has been shown to be very effective in the separation of proteins. All samples were measured at a flow rate of 1 ml / minute and each analysis was completed within 20 minutes.

Example 6

A gel was prepared in the same manner as in Example 1 except that a liquid mixture of 100 g of vinyl acetate / 48 g (X = 0/33) triallyl isocyanurate / 104 g n-heptanol and 3/7 g of 2,2'- Azobisisobutyronitrile used instead of the liquid mixture of Example 1.

The resulting gel had a .Dw of 9> 7 μm, a q _0J , of 4/4 meq / g (an ester exchange ratio of 0/47), a W _R of 1.5 g of water / g dry gel, and a specific active ingredient.

same surface of 60 m / g.

This gel was placed in the same column as in example 1.

- »231105 7

and the analysis of polyethylene glycols and dextrans with different molecular weights was carried out using the packed column. They were eluted in the order of higher molecular weight.

ated _e M ,. for dextran was 5 χ 10 ₀ All samples were measured at a flow rate of 1 ml / minute and each analysis was completed within 20 minutes.

Example 7

A gel was prepared in the same manner as in Example 1! only a liquid mixture of 100 g vinyl acetate, 52 g (X - 0.35) triallyl isocyanurate, 122 g n-butyl acetate, 6 g polyvinyl acetate having a degree of polymerization of about 500 and 3.8 g of 2,2'-azobisisobutyronitrile instead of the liquid Mixtures of Example 1 were used and the ester exchange reaction was carried out at 40 ^° C. for 20 hours.

The resulting gel had a Dw of 11.4 / Um, a q _0} , of 7.6 meq / g (a .75 die exchange ratio), one

Wp of 1.65 g of water / g dry gel and a specific act 2

same surface of 95 m / g.

This gel was filled in the same column as in Example 1, and the analysis of polyethylene glycols and dextranes having different molecular weights and proteins such as bovine serum albumin, ovalbumin and myoglobin as standard samples was carried out using the packed column. As a result, it was confirmed that the substances of each group were eluted in order of the substance having a higher molecular weight and the recovery rate of the proteins was substantially 100 % . The M ₁ - ^ of dextran was calculated as 2 χ 10. All samples were analyzed at a

speed of 1 ml / minute / and each analysis was completed within 20 minutes.

Bg.iep ie 1_8

A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 100 g of vinyl acetate, 64 g (X «0.4) of triallyl isocyanurate, 164 g of n-butyl acetate, 8.2 g of polyvinyl acetate having a degree of polymerization of about 500 and 4.1 g of 2,2'-azobisisobutyronitrile were used in place of the liquid mixtures of Example 1 and the ester exchange reaction was carried out at 40 ^° C. for 20 hours *

The resulting gel had a Dw of 11 / .8 Aim, one of 5.2 meq / g (an ester exchange ratio of 0.6). a W ₀ of 1.85 g of water / g dry gel and a specific active

2 same surface of 120 m / g.

This gel was filled in the same column as in Example 1, and the analysis of polyethylene glycols and dextranes having different molecular weights and proteins such as albumin, ovalbumin and myoglobin was carried out using the packed column. It was confirmed that the substances of each group in the Order of the substance were eluted with a higher molecular weight and that the recovery rate of the proteins was substantially 100 % . The M ,. for dextran was calculated as 1 χ 10. All samples were measured at a flow rate of 1 ml / minute and each analysis was completed within 20 minutes. ,

Example 9

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A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 100 g of vinyl acetate, 37.5 g (X = 0.28) of triallyl isocyanurate, 69 g of n-butyl acetate, 69 g of decalin (a mixture of cis and trans), 4 g polyvinyl be mixed with an degree of polymerization of about 500 and 3.4 g of 2,2'-azobisisobutyronitrile ~ used instead of the liquid Gernischs of example 1 and the Esterauetauschreaktion carried out at 40 ⁰ C for 20 hours.

The resulting gel had a Dw of 14.5 g, a q _Q "of 8.5 meq / g (a Esteraustauschverhältnis of 0.74), a VV ₀ of 1.98 g water / g dry gel and a specific effective

K 2

same surface of 80 m / g _# .

This gel was filled in the same column as in Example 1, and the analysis of polyethylene glycols and dextrans with different molecular weights and of proteins such as α-globulin, ovalbumin and myoglobin as standard samples for protein was carried out using the packed column Substances of each group were eluted in the order of the substance with a higher molecular weight and that the recovery rate of the proteins was substantially 100 % . The M ₁ . for Dextrane bec χ im

carried 1 χ 10 All samples were measured at a flow rate of 1 ml / minute and each analysis was completed within 20 minutes.

Comparative Example 1

A gel was prepared in the same manner as in Example 1, except that the saponification was carried out at 60 ^° C. for 20 hours in one of 60 g of sodium hydroxide, 100 g of water and 500 ml

231105

Methanol solution instead of the ester

, ; is ·

Exchange reaction of Example 1.

The resulting gel had a Dw of 9/7 to give a q ₀ H of 13.5 meq / g (an ester exchange ratio of 0.98), a W "of 1.95 g of water / g dry gel. After the infrared absorption spectrum of the gel, the absorption due to the ester group at 1730 era completely disappeared.

This gel was filled in the same column as in Example 1, and the analysis was carried out under the same chromatographic conditions as in Example 1, but the pressure loss in the column was so increased due to the insufficient mechanical strength of the gel that no measurement was possible was.

A gel was prepared in the same manner as in Example 1 except that the ester exchange reaction was carried out at ⁰ ° C. for 5 hours.

The resulting gel had a Dw of 9.0 / um, a q _o "of 3.2 ra.eq / g (an ester exchange ratio of 0.33) and a VV _R of 1.24 g water / g dry gel.

The gel had a high mechanical strength, and a solvent could be passed through the same column filled with this gel as in Example 1 at a flow rate of 1 liter / minute or more, but the analysis of α-globulin was due to the abnormally large eluted

--- 2311057

Volume unsuccessful / and also the recovery ratio was low.

Comparative Example 3 .

A uniform liquid mixture of 90 g of vinyl acetate / 4 / 5.g (X = 0/017) triallyl isocyanurate, 60 g of toluene and 0.9 g of benzoyl peroxide and 210 g of an aqueous solution containing 2 % of a polyvinyl alcohols having a degree of polymerization of 500 and a saponification ratio of 89 % / were filled in a 1-1 flask, and the suspension polymerization was carried out at 60 ⁰ C for 16 hours. The resulting polymer particles were separated by filtration, washed with water and acetone, dried and then saponified at 60 ^° C. for 30 hours in a liquid mixture of 50 g sodium hydroxide, 100 g methanol and 100 g water.

The resulting gel had a Dw of 14.8 μm, a q _0H of 18.2 meq / g (a saponification ratio of 0.90), a VVp of 3/31 g of water / g dry gel and a specific effective surface area of 0 , 1 m / g, suggesting that there would hardly be a pore in the gel when dry.

This gel was filled in the same column as in Example 1 and the analysis was carried out under the same chromatographic conditions as in Example 1, but due to the insufficient mechanical strength of the gel, the pressure loss in the column was increased so that no measurement was possible was.

231 1 0 5 7

A gel was prepared in the same manner as in Example 1, except that a liquid mixture of 50 g of vinyl acetate / 73 g. (X - 0.6) triallyl isocyanurate, 123 g n-butyl acetate and 3 g of 2,2 '' ° azobisisobutyronitrile used in place of the liquid mixtures of Example 1 and the Esteraustauschreaktion for 20 hours at 40 ⁰ C. "

The resulting gel had a Dw of 9.2, at, a of 3.3 tn eq / g (an ester exchange ratio of 0.61) and a W _R of 1.4 g water / g dry gel *

This gel was filled in the same column as in Example 1, and the analysis of bovine serum albumin, ovalbumin and myoglobin as standard samples was carried out, but all were adsorbed on the gel, so that almost no elution was carried out and no peak could be identified .

Comparative Example 5

It was prepared as in Example 1 a gel in 6 same manner, but was a liquid Geraisch of 100 g of vinyl acetate, 11 g (X - 0.1) triallyl isocyanurate, 44 g of toluene and 2.8 g of 2,2'-azobisisobutyronitrile instead of of the liquid mixture of Example 1 is used.

The resulting gel had a Dw of 10.1 μm, a q _0H of 12.5 Ri.eq / g (an ester exchange ratio of 0.78) and a W _R of 1.97 g water / g dry gel,

2 3110 5 7

This gel was filled in the same column as in Example 1 and the analysis was carried out under the same chromatographic conditions as in Example 1, but the insufficient mechanical strength of the gel increased the pressure drop in the column. that no measurement could be made.

While the invention has been described in detail with reference to specific embodiments, it will be obvious to those skilled in the art that various changes and modifications are possible without departing from its spirit and scope.

Claims (24)

-ZZ GZ 14588 57 E rf in η g sap sprue h; 1, completely porous filling material for the fast fluid «. Chromatography, characterized in that it comprises a granular cross-linked copolyester which consists essentially of (I) units of at least one vinyl alcohol, (II) units of at least one vinyl ester of a carboxylic acid and (III) units of at least one cross-linking monomer with an isocyanurate ring, wherein the ratio of aer units (I) to units (II) in the copolyring is within the range corresponding to the following equation: about 0.4 pounds a ~ f ~ 5 ^ about 0.8 in which · · ' a_ and Id molar ratios of units (-1)
or (II) in the total units (I), (II)
and (III) the copolymer. 2 3110 5 7 2. Filling material according to item 1, characterized in that the ratio of the units (I) to the units (II) in the copolymer within the following equation , speaking area lies: about 0.45 ^. · ^ Ι ^ k about 0.75 3 5 glycol or dextran in the range of about 10 to about 10 lies. , , · 3. ^ filler material according to item 1, characterized in that the ratio of units (I), (II) and (III) mers .in the copoly "is within the aer following equation corresponding region: about 0.20 ^ - ^ ' ³ g- ^ about 0.40 a_ and b_ have the same meaning as above, and c_ the molar ratio of the units (III)
in the total units (I), (II) and (III) of the copolymer. " 231105 7 4. Filler material according to item 1 or 3, characterized in that the units of the vinyl ester of a carboxylic acid are those of the vinyl ester of a carboxylic acid having 2 to 5 carbon atoms. 5. filler according to item 4, characterized in that the vinyl ester of the carboxylic acid is selected from the group comprising vinyl acetate, vinyl n-propionate, vinyl butyrate, vinyl valerate and vinyl pivalate. 6. filler according to item 5, characterized in that the vinyl ester of the carboxylic acid is vinyl acetate. 7. filler according to item 5, characterized in that it is vinyl propionate in the vinyl ester of the carboxylic acid. 8. Filler material according to item 1 or 3, characterized in that the units of the crosslinking monomer having an isocyanurate ring are those of the crosslinking monomer having the following formula: R ₁ , R ₂ and R ₂ may be the same or different and are selected from the group consisting of -CH ₂ -CH = CH ₂ , -CH-CsCH and -CH ₂ -C = CH ₂ are selected. 9 «filling material according to item 8, characterized in that it is the crosslinking monoineren triallylisocyanurate. 10. Filling material according to item 3, characterized in that the ratio of units (1), (II) and (III) is within the range corresponding to the following equation: about 0; 24 ^ _A £ about 0.32 - a + υ + each wherein £ »k ^unc * £ have the meaning explained above, and the exclusive molecular weight of polyethylene 11. Filling material according to item 3, characterized in that the ratio of units (I), (II) and (III) is in the range corresponding to the following equation: about 0.27? =, - - about 0.35 Oz wherein a, b, and q have the same meaning as explained above, and the exclusive molecular weight of polyethylene glycol and dextran ranges from about 10 to about 10. "_.". · 12, filler according to item 1, characterized in that it has a water recovery of about 0.5 to about 2.0 g of water / g dry gel. ' 13. Filling material according to item 12, characterized in that the IVasserrückgewinnüng is in the range of about 0.8 to about 2.0 g of water / g dry gel. 14. A filler according to item 1, characterized in that the copolymer has a weight-average particle diameter of about 5 to 20 yUm 4 15. A filler according to item 14, characterized in that the copolymer has a weight average particle diameter of about 5 to 12 ^ to. 16. A process for the preparation of a filling material for rapid liquid chromatography, characterized in that a monomer mixture of at least one vinyl ester of a carboxylic acid having 2 to 5 carbon atoms and at least one cross-linking monoenera of the formula: ^R 3 11 in which R ₁ , Rp and R ^ may be the same or different and are selected from the group consisting of -.CH ₂ -CH-CH ₂ , -CH-C = CH and -CHp-C-Cl-L * are selected, oer subjected to suspension polymerization, and the resulting granular copolymers of the Esteraustauschreaktion or the saponification is up to Einein such Grado subjected that the Esteraustauschverhältnis or the saponification ratio is about 0.4 to about 0.8. 17. The method according to item 16, characterized in that the Ester exchange ratio or saponification ratio u is about 0.45 to about 0.75. 18. A method according to item 16, characterized in that the molar ratio of the vinyl ester of the carboxylic acid and the crosslinking monomer 1 is from about 0.08 to about 0.22. 19. A method according to item 18, characterized in that the molar ratio of the vinyl ester of the carboxylic acid and the crosslinking monomer 1 is from about 0.11 to about 0.16. 20. The method of item 13, characterized in that the molar ratio of the vinyl ester of the carboxylic acid and the crosslinking monomer is 1 to about 0.12 to about 0.18. · 231105 7 21. The method according to item 16, characterized in that the vinyl ester of the carboxylic acid from the vinyl acetate, vinyl propionate, vinyl butyrate, Vinylvalerät and Vinylpivalat comprehensive group is selected. 22. A method according to item 21, characterized in that the vinyl ester of the carboxylic acid is vinyl acetate. 23. The method according to item 21, characterized in that it is vinyl propionate in the vinyl ester of the carboxylic acid. 24. The method according to item 16 ', characterized in that the crosslinking inonoin is triallylisocyanurate.